五藤定神汤减轻多巴胺能神经元丢失并抑制MPTP模型M1型小胶质细胞极化的研究*

作者：吴幸鸿1，梁慧荟2，戴 健1，韦秀英1，李若琪1，陈钊森1，王凯华2，王 荔1，2

单位：1.广西中医药大学，广西 南宁 530200； 2.广西中医药大学附属国际壮医医院，广西 南宁 530201

引用：引用：吴幸鸿，梁慧荟，戴健，韦秀英，李若琪，陈钊森，王凯华，王荔.五藤定神汤减轻多巴胺能神经元丢失并抑制MPTP模型M1型小胶质细胞极化的研究[J].中医药导报,2026,32(1):1-6,28.

DOI：10.13862/j.cn43-1446/r.2026.01.001

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摘要：

目的：研究五藤定神汤是否具有改善多巴胺（DA）能神经元丢失和调控MPTP模型M1小胶质细胞活化的作用。方法：将36只C57BL/6J小鼠随机分为空白组、模型组和干预组，每组12只。采用腹腔注射MPTP方法构建帕金森病（PD）小鼠模型。干预组小鼠灌服相应剂量的五藤定神汤，空白组、模型组灌服超纯水，1次/d，连续14 d。于实验第0天、第5天、第19天对各组小鼠进行行为学检测，实验第20天处死小鼠，采用免疫荧光染色对小鼠中脑黑质进行酪氨酸羟化酶（TH）、小胶质细胞离子钙结合衔接分子1（Iba-1）、一氧化氮合酶（iNOS）染色，采用高效液相色谱检测纹状体多巴胺含量，RT-qPCR检测纹状体Iba mRNA、iNOS mRNA表达水平，蛋白质印迹（Western blotting）法检测黑质Iba、iNOS蛋白表达水平，酶联免疫吸附试验（ELISA）检测纹状体肿瘤坏死因子-α（TNF-α）、白介素-1（IL-1）的含量。结果：与模型组相比，干预组爬杆实验时长显著缩短，旷场实验路程增加、平均移动速度均显著提高（P＜0.01），黑质络氨酸羟化酶表达明显增加，纹状体DA含量显著提高（P＜0.05）。此外，相对模型组，免疫荧光染色、WB、RT-qPCR检测均提示干预组iNOS较模型组减少（P＜0.05）；ELISA检测提示TNF-α、IL-1β含量减少。结论：五藤定神汤可改善MPTP模型小鼠运动障碍症状，抑制小胶质细胞M1型活化，降低纹状体TNF-α、IL-1β水平，减轻中脑黑质致密部多巴胺能神经元丢失。

关键词：帕金森病；五藤定神汤；小胶质细胞；M1型极化；多巴胺能神经元；小鼠

Abstract：

Objective: To investigate whether Wuteng Dingshen decoction can ameliorate the loss of dopaminergic neurons and regulate the activation of M1 microglia in MPTP-induced Parkinson's disease (PD) mouse models. Methods: A total of 36 C57BL/6J mice were randomly divided into a blank group, a model group, and an intervention group, 12 mice in each. PD mouse models were established by intraperitoneal injection of MPTP. The intervention group was intragastrically administered with Wuteng Dingshen decoction, while the blank group and the model group received ultrapure water, once daily for 14 consecutive days. Behavioral tests were performed on day 0, 5, and 19. All mice were sacrificed on day 20. Immunofluorescence staining was used to detect tyrosine hydroxylase (TH), ionized calcium-binding adapter molecule 1 (Iba-1), and inducible nitric oxide synthase (iNOS) in the substantia nigra. High-performance liquid chromatography was used to measure dopamine (DA) content in the striatum. RT-qPCR was used to detect the  levels of Iba-1 mRNA and iNOS mRNA in the striatum. Western blotting was used to determine the protein expression levels of Iba-1 and iNOS in the substantia nigra. Enzyme-linked immunoadsordent assay (ELISA) was used to measure the levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in the striatum. Results: Compared with the model group, the intervention group showed a significantly shortened duration in the pole-climbing test, and increased total distance and average speed in the open-field test (P<0.01). TH expression in the substantia nigra and DA content in the striatum were significantly higher in the intervention group (P<0.05). Furthermore, immunofluorescence staining, Western blotting, and RT-qPCR analyses all indicated a reduction in iNOS in the intervention group compared to the model group (P<0.05). ELISA revealed decreased levels of TNF-α and IL-1β. Conclusion: Wuteng Dingshen decoction can ameliorate motor dysfunction, inhibit M1 microglial polarization, reduce the levels of TNF-α and IL-1β, and attenuate the loss of dopaminergic neurons in the substantia nigra pars compacta of MPTP-induced PD mice.

Key words：Parkinson's disease; Wuteng Dingshen decoction; microglia; M1 polarization; dopaminergic neurons; mice

发布时间：2026-01-30

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